Phytochemical and Biological Evaluation and Isolation of Quercetin from Hypericum cordifolium Flower

M. Rajbhandari, Janak Khatri, Gitanjali Gautam, Kusum Maiya Dhoubhadel
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Abstract

Hypericum cordifolium is a medicinal plant used in the treatment of various ailments in Nepalese traditional medicine. The medicinal values and bioactive constituents of Hypericum perforatum have been well investigated and is used as an alternative medicine in the treatment of depression, cancer, inflammation, bacterial and viral infections. However, scientific reports on Nepalese H. cordifolium are very limited. Here we report about the chemical screening of extracts, quality analysis of H. cordifolium extract by thin layer chromatography, TLC using H. perforatum extract and hyperoside as the standards, estimation of phenolics, flavonoids, tannins and sugars as well as antioxidant, antibacterial and cytotoxic activities of flower extracts. The ethyl acetate extract was fractionated and some phytochemicals were isolated. In our findings, TLC analysis of H. cordifolium extract showed the presence of chlorogenic acid, hyperoside and quercetin. Hypericine and rutin were not detected which indicated that the chemotaxonomy of H. cordifolium is different from H. perforatum. In phytochemical analysis, ethyl acetate extract showed the greater amounts of flavonoids, methanol extract showed the greater amounts of tannins and sugars and 50% aqueous methanol extract showed the greater amounts of phenolics. In DPPH free radical scavenging assay, ethyl acetate (18.79±0.98 μg/ml) and 50% aq. methanol (18.98±0.81 μg/ml) extracts showed the lower IC50 values than the methanol extract (27.00±1.03 μg/ml) due to the presence of high amounts of flavonoids and phenolics in these extracts. In antibacterial assay, all extracts showed moderate antibacterial activity against Staphylococcus aureus except 50% aqueous methanol extract. In cytotoxicity assay on human bladder carcinoma cell line 5637, ethanol extract showed IC50 value of 41.35±5.01 μg/ ml. Quercetin was isolated from ethyl acetate extract and characterized by UV and FTIR spectroscopy. The present findings indicated that the flowers of H. cordifolium could be the rich source bioactive compounds with antibacterial, cytotoxic and antioxidant activities.
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金丝桃花中槲皮素的植物化学生物学评价及分离
金丝桃(Hypericum cordifolium)是尼泊尔传统医学中用于治疗各种疾病的药用植物。贯叶连翘的药用价值和生物活性成分已被充分研究,并被用作治疗抑郁症、癌症、炎症、细菌和病毒感染的替代药物。然而,关于尼泊尔堇青花的科学报道非常有限。本文报道了提取物的化学筛选、贯叶连翘提取物的薄层色谱质量分析、以贯叶连翘提取物和金丝桃苷为标准的薄层色谱分析、花提取物的酚类、黄酮类、单宁类和糖的含量以及抗氧化、抗菌和细胞毒活性的测定。对乙酸乙酯提取物进行了分馏,分离出了部分植物化学物质。在我们的研究中,薄层色谱分析显示,堇叶提取物中含有绿原酸、金丝桃苷和槲皮素。未检出金丝桃素和芦丁,表明其化学分类与贯叶连翘不同。在植物化学分析中,乙酸乙酯提取物中黄酮类化合物含量较高,甲醇提取物中单宁和糖含量较高,50%甲醇水溶液提取物中酚类物质含量较高。在DPPH自由基清除实验中,乙酸乙酯(18.79±0.98 μg/ml)和50%甲醇(18.98±0.81 μg/ml)提取物的IC50值低于甲醇提取物(27.00±1.03 μg/ml),这是由于乙酸乙酯(18.79±0.98 μg/ml)提取物中含有大量的黄酮类和酚类物质。抑菌试验表明,除50%甲醇水溶液提取物外,其余提取物对金黄色葡萄球菌均有中等抑菌活性。在对人膀胱癌细胞株5637的细胞毒性实验中,乙醇提取物的IC50值为41.35±5.01 μg/ ml。槲皮素从乙酸乙酯提取物中分离得到,并用紫外光谱和红外光谱对槲皮素进行了表征。研究结果表明,堇青花可能是具有抗菌、细胞毒和抗氧化活性的生物活性物质的丰富来源。
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